The monoclonal antibody Kat5-18 recognizes T-cell immunoglobulin and mucin domain containing molecule (TIMD-4, also called TIM-4). TIMD-4 is a type I transmembrane protein belonging to the immunoglobulin superfamily, containing one Ig-like V-type (immunoglobulin-like) domain. TIMD-4 is expressed in Mac1+ cells in spleen, lymph nodes and fetal liver. TIMD-4 binds apoptotic cells by recognizing phosphatidylserine via its immunoglobulin domain. TIMD-4 in fibroblasts enhanced their ability to engulf apoptotic cells. TIMD-4 is involved in regulating T-cell proliferation and lymphotoxin signaling. TIMD-4 and TIMD-1 are phosphatidylserine receptors for the engulfment of apoptotic cells, and are also involved in intracellular signaling in which exosomes are involved.
The monoclonal antibody Kat5-18 blocks the engulfment of apoptotic cells by thymic macrophages.
Flow cytometry, Functional studies, Immuno assays, Immuno fluorescence
FC: Cells were incubated on ice for 30 min with 1 μg/ml biotinylated Kat5-18 in 100 μl staining solution (PBS containing 2% FCS), followed by incubation with 2 μg/ml Allophycocyanin (APC)-Cyanine 7 (Cy7)-conjugated streptavidin and 2 μg/ml APCconjugated anti-CD11b. The cells were then stained with 0.5 μM SYTOX Blue (Ref.2).
FS: Antibody Kat5-18 blocks the engulfment of apoptotic cells by thymic macrophages (Ref.1).
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.